Maldonado Martin, Huang Tianhua, Chen Jianying, Zhong Ying
Reproductive Medicine & Genetics, Chengdu Jinjiang Hospital for Maternal & Child Health Care, Chengdu 610066, China.
Stem Cells Int. 2018 Oct 11;2018:7609284. doi: 10.1155/2018/7609284. eCollection 2018.
In vitro maturation (IVM) in cumulus oocyte complexes (COCs) can be improved by the presence of human Wharton's jelly-derived MSCs (hWJ-MSCs), under specific culture conditions. COCs were cultured in twelve different culture systems, composed of four stock media, stock media conditioned with hWJ-MSCs, and stock media in which the oocytes were indirectly cocultured with the hWJ-MSCs. The rates of maturation to meiosis II were compared among the groups. G2-PLUS and coculture with DMEM-F12 were the most efficient systems for the maturation of COCs. The fertilization rate and rate of development to the blastocyst stage were compared between these two groups. Moreover, hWJ-MSC-conditioned media showed no benefits for the COC-IVM. The analysis of OCT4 expression of hWJ-MSCs in G1-PLUS, TYH, and G2-PLUS showed a downregulation of OCT4 by 25.9, 24.7, and 6.6%, respectively, compared to that in hWJ-MSCs cultured in DMEM-F12. Finally, we have demonstrated that two prerequisites appeared to be necessary for the hWJ-MSCs to improve the IVM of COCs: hWJ-MSCs' differentiation potential and the presence of coordinated paracrine interaction between the stem cells and COCs. Under the appropriate conditions, the paracrine factors produced in the coculture system with DMEM-F12 may help to develop synthetic media for successful in vitro culture of COCs.
在特定培养条件下,人脐带华通氏胶间充质干细胞(hWJ-MSCs)的存在可改善卵丘卵母细胞复合体(COCs)的体外成熟(IVM)。将COCs培养于12种不同的培养体系中,这些体系由4种基础培养基、用hWJ-MSCs处理过的基础培养基以及使卵母细胞与hWJ-MSCs间接共培养的基础培养基组成。比较各实验组中减数分裂II期的成熟率。G2-PLUS培养基以及与DMEM-F12共培养是COCs成熟的最有效体系。比较这两组之间的受精率和囊胚期发育率。此外,hWJ-MSC条件培养基对COC-IVM并无益处。对G1-PLUS、TYH和G2-PLUS培养基中hWJ-MSCs的OCT4表达分析显示,与在DMEM-F12中培养的hWJ-MSCs相比,OCT4分别下调了25.9%、24.7%和6.6%。最后,我们证明hWJ-MSCs改善COCs的IVM似乎有两个必要前提条件:hWJ-MSCs的分化潜能以及干细胞与COCs之间存在协同旁分泌相互作用。在适当条件下,与DMEM-F12共培养体系中产生的旁分泌因子可能有助于开发用于COCs成功体外培养的合成培养基。